Systems Biology, School for Marine Science and Technology, Zhejiang Ocean University, Zhoushan 316022, China.
Genes (Basel). 2024 May 7;15(5):590. doi: 10.3390/genes15050590.
Pa0665 in shares homologous sequences with that of the essential A-type iron-sulfur (Fe-S) cluster insertion protein ErpA in . However, its essentiality in and its complementation with has not been experimentally examined. To fulfill this task, we constructed plasmid-based -mutant / using a three-step protocol. The mutant displayed growth defects at 42 °C, which were complemented by expressing . Microscopic observations indicated a petite cell phenotype for / at 42 °C, correlated with the downregulation of the gene. RNA sequencing revealed significant transcriptional changes in genes associated with the oxidative phosphorylation (OXPHOS) system, aligning with reduced ATP levels in / under 42 °C. Additionally, the -mutant showed heightened sensitivity to HO at 42 °C. Overall, our study demonstrates the essential role of for OXPHOS function and is complemented by . We propose that the plasmid-based -allele is useful for genetic analysis of essential genes of interest in .
Pa0665 在与必需的 A 型铁硫 (Fe-S) 簇插入蛋白 ErpA 的同源序列方面具有同源性。然而，它在 中的必要性及其与 的互补性尚未通过实验检验。为了完成这项任务，我们使用三步方案构建了基于质粒的 / 突变体。该突变体在 42°C 时表现出生长缺陷，这可以通过表达 来弥补。显微镜观察表明 / 在 42°C 时表现出小细胞表型，与 基因的下调相关。RNA 测序显示与氧化磷酸化 (OXPHOS) 系统相关的基因发生了显著的转录变化，与 42°C 下的 ATP 水平降低一致。此外，-突变体在 42°C 时对 HO 表现出更高的敏感性。总的来说，我们的研究表明 对于 OXPHOS 功能是必需的，并且可以由 来补充。我们提出基于质粒的 -等位基因可用于研究 在感兴趣的必需基因的遗传分析。
